Team:Glendale CC AZ/Protocols/Ligation

From 2013.igem.org

(Difference between revisions)
(→Procedure)
 
(5 intermediate revisions not shown)
Line 1: Line 1:
-
{{:Team:Glendale_CC_AZ/main.css}}
+
{{:Team:Glendale_CC_AZ/main.css}}  
 +
<html>
 +
<head>
 +
<link
 +
href="http://fonts.googleapis.com/css?family=PT+Serif:400,700,400italic,700italic"
 +
rel="stylesheet" type="text/css">
 +
<link
 +
href="http://fonts.googleapis.com/css?family=Source+Sans+Pro:300,700,300italic"
 +
rel="stylesheet" type="text/css">
 +
<meta charset="UTF-8">
 +
<style type="text/css">
 +
body {
 +
  margin: 0;
 +
  padding: 0;
 +
  background: rgb(230,230,230);
 +
 
 +
  color: rgb(50,50,50);
 +
  font-family: 'Open Sans', sans-serif;
 +
  font-size: 112.5%;
 +
  line-height: 1.6em;
 +
}
-
== Ligation Protocol ==
+
/* ================ The Timeline ================ */
 +
 
 +
.timeline {
 +
  position: relative;
 +
  width: 660px;
 +
  margin: 0 auto;
 +
  margin-top: 20px;
 +
  padding: 1em 0;
 +
  list-style-type: none;
 +
}
 +
p list {
 +
  text-align: left;
 +
}
 +
 
 +
.timeline:before {
 +
  position: absolute;
 +
  left: 50%;
 +
  top: 0;
 +
  content: ' ';
 +
  display: block;
 +
  width: 6px;
 +
  height: 100%;
 +
  margin-left: -3px;
 +
  background: rgb(80,80,80);
 +
  background: -moz-linear-gradient(top, rgba(80,80,80,0) 0%, rgb(80,80,80) 8%, rgb(80,80,80) 92%, rgba(80,80,80,0) 100%);
 +
  background: -webkit-gradient(linear, left top, left bottom, color-stop(0%,rgba(30,87,153,1)), color-stop(100%,rgba(125,185,232,1)));
 +
  background: -webkit-linear-gradient(top, rgba(80,80,80,0) 0%, rgb(80,80,80) 8%, rgb(80,80,80) 92%, rgba(80,80,80,0) 100%);
 +
  background: -o-linear-gradient(top, rgba(80,80,80,0) 0%, rgb(80,80,80) 8%, rgb(80,80,80) 92%, rgba(80,80,80,0) 100%);
 +
  background: -ms-linear-gradient(top, rgba(80,80,80,0) 0%, rgb(80,80,80) 8%, rgb(80,80,80) 92%, rgba(80,80,80,0) 100%);
 +
  background: linear-gradient(to bottom, rgba(80,80,80,0) 0%, rgb(80,80,80) 8%, rgb(80,80,80) 92%, rgba(80,80,80,0) 100%);
 +
 
 +
  z-index: 5;
 +
}
 +
 
 +
.timeline li {
 +
  padding: 1em 0;
 +
}
 +
 
 +
.timeline li:after {
 +
  content: "";
 +
  display: block;
 +
  height: 0;
 +
  clear: both;
 +
  visibility: hidden;
 +
}
 +
 
 +
.direction-l {
 +
  position: relative;
 +
  width: 300px;
 +
  float: left;
 +
  text-align: right;
 +
}
 +
 
 +
.direction-r {
 +
  position: relative;
 +
  width: 300px;
 +
  float: right;
 +
}
 +
 
 +
.flag-wrapper {
 +
  position: relative;
 +
  display: inline-block;
 +
 
 +
  text-align: center;
 +
}
 +
 
 +
.flag {
 +
  position: relative;
 +
  display: inline;
 +
  background: rgb(248,248,248);
 +
  padding: 6px 10px;
 +
  border-radius: 5px;
 +
 
 +
  font-weight: 600;
 +
  text-align: left;
 +
}
 +
 
 +
.direction-l .flag {
 +
  -webkit-box-shadow: -1px 1px 1px rgba(0,0,0,0.15), 0 0 1px rgba(0,0,0,0.15);
 +
  -moz-box-shadow: -1px 1px 1px rgba(0,0,0,0.15), 0 0 1px rgba(0,0,0,0.15);
 +
  box-shadow: -1px 1px 1px rgba(0,0,0,0.15), 0 0 1px rgba(0,0,0,0.15);
 +
}
 +
 
 +
.direction-r .flag {
 +
  -webkit-box-shadow: 1px 1px 1px rgba(0,0,0,0.15), 0 0 1px rgba(0,0,0,0.15);
 +
  -moz-box-shadow: 1px 1px 1px rgba(0,0,0,0.15), 0 0 1px rgba(0,0,0,0.15);
 +
  box-shadow: 1px 1px 1px rgba(0,0,0,0.15), 0 0 1px rgba(0,0,0,0.15);
 +
}
 +
 
 +
.direction-l .flag:before,
 +
.direction-r .flag:before {
 +
  position: absolute;
 +
  top: 50%;
 +
  right: -40px;
 +
  content: ' ';
 +
  display: block;
 +
  width: 12px;
 +
  height: 12px;
 +
  margin-top: -10px;
 +
  background: #fff;
 +
  border-radius: 10px;
 +
  border: 4px solid rgb(255,80,80);
 +
  z-index: 10;
 +
}
 +
 
 +
.direction-r .flag:before {
 +
  left: -40px;
 +
}
 +
 
 +
.direction-l .flag:after {
 +
  content: "";
 +
  position: absolute;
 +
  left: 100%;
 +
  top: 50%;
 +
  height: 0;
 +
  width: 0;
 +
  margin-top: -8px;
 +
  border: solid transparent;
 +
  border-left-color: rgb(248,248,248);
 +
  border-width: 8px;
 +
  pointer-events: none;
 +
}
 +
 
 +
.direction-r .flag:after {
 +
  content: "";
 +
  position: absolute;
 +
  right: 100%;
 +
  top: 50%;
 +
  height: 0;
 +
  width: 0;
 +
  margin-top: -8px;
 +
  border: solid transparent;
 +
  border-right-color: rgb(248,248,248);
 +
  border-width: 8px;
 +
  pointer-events: none;
 +
}
 +
 
 +
.time-wrapper {
 +
  display: inline;
 +
 
 +
  line-height: 1em;
 +
  font-size: 0.66666em;
 +
  color: rgb(250,80,80);
 +
  vertical-align: middle;
 +
}
 +
 
 +
.direction-l .time-wrapper {
 +
  float: left;
 +
}
 +
 
 +
.direction-r .time-wrapper {
 +
  float: right;
 +
}
 +
 
 +
.time {
 +
  display: inline-block;
 +
  padding: 4px 6px;
 +
  background: rgb(248,248,248);
 +
}
 +
 
 +
.desc {
 +
  margin: 1em 0.75em 0 0;
 +
 
 +
  font-size: 0.77777em;
 +
  font-style: italic;
 +
  line-height: 1.5em;
 +
}
 +
 
 +
.direction-r .desc {
 +
  margin: 1em 0 0 0.75em;
 +
}
 +
 
 +
/* ================ Timeline Media Queries ================ */
 +
 
 +
@media screen and (max-width: 660px) {
 +
 
 +
.timeline {
 +
width: 100%;
 +
padding: 4em 0 1em 0;
 +
}
 +
 
 +
.timeline li {
 +
padding: 2em 0;
 +
}
 +
 
 +
.direction-l,
 +
.direction-r {
 +
float: none;
 +
width: 100%;
 +
 
 +
text-align: center;
 +
}
 +
 
 +
.flag-wrapper {
 +
text-align: center;
 +
}
 +
 
 +
.flag {
 +
background: rgb(255,255,255);
 +
z-index: 15;
 +
}
 +
 
 +
.direction-l .flag:before,
 +
.direction-r .flag:before {
 +
  position: absolute;
 +
 
 +
  top: -30px;
 +
left: 50%;
 +
content: ' ';
 +
display: block;
 +
width: 12px;
 +
height: 12px;
 +
margin-left: -9px;
 +
background: #fff;
 +
border-radius: 10px;
 +
border: 4px solid rgb(255,80,80);
 +
z-index: 10;
 +
}
 +
 
 +
.direction-l .flag:after,
 +
.direction-r .flag:after {
 +
content: "";
 +
position: absolute;
 +
left: 50%;
 +
top: -8px;
 +
height: 0;
 +
width: 0;
 +
margin-left: -8px;
 +
border: solid transparent;
 +
border-bottom-color: rgb(255,255,255);
 +
border-width: 8px;
 +
pointer-events: none;
 +
}
 +
 
 +
.time-wrapper {
 +
display: block;
 +
position: relative;
 +
margin: 4px 0 0 0;
 +
z-index: 14;
 +
}
 +
 
 +
.direction-l .time-wrapper {
 +
float: none;
 +
}
 +
 
 +
.direction-r .time-wrapper {
 +
float: none;
 +
}
 +
 
 +
.desc {
 +
position: relative;
 +
margin: 1em 0 0 0;
 +
padding: 1em;
 +
background: rgb(245,245,245);
 +
-webkit-box-shadow: 0 0 1px rgba(0,0,0,0.20);
 +
-moz-box-shadow: 0 0 1px rgba(0,0,0,0.20);
 +
box-shadow: 0 0 1px rgba(0,0,0,0.20);
 +
 +
  z-index: 15;
 +
}
 +
 
 +
.direction-l .desc,
 +
.direction-r .desc {
 +
position: relative;
 +
margin: 1em 1em 0 1em;
 +
padding: 1em;
 +
 +
  z-index: 15;
 +
}
 +
 
 +
}
 +
 
 +
@media screen and (min-width: 400px ?? max-width: 660px) {
 +
 
 +
.direction-l .desc,
 +
.direction-r .desc {
 +
margin: 1em 4em 0 4em;
 +
}
 +
 
 +
} 
 +
* {
 +
-webkit-box-sizing: border-box;
 +
-moz-box-sizing: border-box;
 +
box-sizing: border-box;
 +
}
 +
 
 +
html {
 +
font-size:50%;
 +
-webkit-font-smoothing: antialiased;
 +
}
 +
 
 +
body {
 +
font-family: "Source Sans Pro";
 +
font-size:1.4rem;
 +
line-height: 2.3rem;
 +
color:#444;
 +
background: #fff;
 +
margin:0;
 +
}
 +
 
 +
a {
 +
color: #06c;
 +
transition: color .3s;
 +
}
 +
 
 +
a:hover {
 +
color: #012c57;
 +
}
 +
 
 +
h1, h2, h3, h4, h5, h6, h7 {
 +
font-family: "Futura";
 +
line-height:2.6rem;
 +
font-weight:normal;
 +
color: #111;
 +
}
 +
 
 +
.content-area {
 +
width:850px;
 +
padding:2rem;
 +
margin:auto;
 +
background: #f9f9f9;
 +
}
 +
 
 +
.content-area h1 {
 +
font-size: 2.7rem;
 +
}
 +
 
 +
.content-area h2 {
 +
font-size: 2.2rem;
 +
}
 +
 
 +
.content-area h3 {
 +
font-size: 2.7rem;
 +
}
 +
 
 +
.content-area h4 {
 +
font-size: 4.0rem;
 +
}
 +
 
 +
.content-area h5 {
 +
font-size: 1.4rem;
 +
}
 +
 
 +
.content-area h6 {
 +
font-size: 1.2rem;
 +
}
 +
 
 +
.content-area aside {
 +
float:right;
 +
width:50%;
 +
padding:2rem;
 +
 
 +
.blockquote {
 +
font-family: Georgia, serif;
 +
font-size: 18px;
 +
font-style: italic;
 +
width: 500px;
 +
margin: 0.25em 0;
 +
padding: 0.35em 40px;
 +
line-height: 1.45;
 +
position: relative;
 +
color: #383838;
 +
}
 +
 
 +
.blockquote:before {
 +
display: block;
 +
padding-left: 10px;
 +
content: "\201C";
 +
font-size: 80px;
 +
position: absolute;
 +
left: -20px;
 +
top: -20px;
 +
color: #7a7a7a;
 +
}
 +
 
 +
.blockquote cite {
 +
color: #999999;
 +
font-size: 14px;
 +
display: block;
 +
margin-top: 5px;
 +
}
 +
 
 +
.blockquote cite:before {
 +
content: "\2014 \2009";
 +
}
 +
 
 +
 
 +
</style>
 +
<title></title>
 +
</head>
 +
<body>
 +
<div class="content-area"> <article>
 +
</article>
 +
<img style="border: 0px solid ; width: 50px; height: 40px;" alt="iGEM" src="http://s21.postimg.org/ff5nkjy9v/IGEM_basic_Logo_stylized.png" align="left">
 +
<h4>Glendale Community College Arizona<img
 +
style="width: 200px; height: 58px;" alt="GCC"
 +
src="https://static.igem.org/mediawiki/2013/f/f1/Gcclogo.gif" align="right">
 +
</h4>
 +
 +
 
 +
<link href='http://fonts.googleapis.com/css?family=Open+Sans:400,300,300italic,400italic,600,600italic,700,700italic' rel='stylesheet' type='text/css'>
 +
 
 +
 
 +
  <h2><p>Protocols</p></h2><img style="border: 0px solid ; width: 400px; height: 200px;" alt="iGEM" src="https://static.igem.org/mediawiki/igem.org/1/19/Tubes2GCC.JPG" align="right">
 +
 
 +
<a href="https://2013.igem.org/Team:Glendale_CC_AZ/Protocols/GrowthCurve">Growth Curve  Assay</a></p>
 +
<a href="https://2013.igem.org/Team:Glendale_CC_AZ/Protocols/NaCl">NaCl Growth Curve Assay</a></p>
 +
<a href="https://2013.igem.org/Team:Glendale_CC_AZ/Protocols/SurivalGrowth">Survival Growth  Assay</a></p>
 +
<a href="https://2013.igem.org/Team:Glendale_CC_AZ/Protocols/AlkalineLysis">Alkaline Lysis Plasmid Miniprep </a></p>
 +
<a href="https://2013.igem.org/Team:Glendale_CC_AZ/Protocols/RestrictionDigest">Restriction Digest</a></p>
 +
<a href="https://2013.igem.org/Team:Glendale_CC_AZ/Protocols/DNAIsolation">DNA Isolation</a></p>
 +
<a href="https://2013.igem.org/Team:Glendale_CC_AZ/Protocols/Bioinformatics">Bioinformatics</a></p>
 +
<a href="https://2013.igem.org/Team:Glendale_CC_AZ/Protocols/Ligation">Ligation</a></p>
 +
<a href="https://2013.igem.org/Team:Glendale_CC_AZ/Protocols/Transformation">Transformation</a></p>
 +
 
 +
 
 +
<!--/.content-area-->
 +
</body>
 +
</html>
 +
 
 +
==== Ligation Protocol ====
Protocol from: http://parts.igem.org/Help:Protocols/Ligation
Protocol from: http://parts.igem.org/Help:Protocols/Ligation
-
== Materials ==
+
==== Materials ====
   -digested plasmid backbone
   -digested plasmid backbone
-
   -equimolar amount of EcoRI-HF SpeI digested fragment
+
   -equimolar amount digested Part 1
-
   -equimolar amount of XbaI PstI digested fragment
+
   -equimolar amount digested Part 2
   -T4 DNA ligase buffer
   -T4 DNA ligase buffer
   -T4 DNA ligase
   -T4 DNA ligase
   -dH2O
   -dH2O
 +
  -thermal cycler
-
== Procedure ==
+
==== Procedure ====
-
 
+
1. Add 2µL of digested plasmid backbone
1. Add 2µL of digested plasmid backbone

Latest revision as of 02:36, 28 September 2013

iGEM

Glendale Community College ArizonaGCC

Protocols

iGEM Growth Curve Assay

NaCl Growth Curve Assay

Survival Growth Assay

Alkaline Lysis Plasmid Miniprep

Restriction Digest

DNA Isolation

Bioinformatics

Ligation

Transformation

Ligation Protocol

Protocol from: http://parts.igem.org/Help:Protocols/Ligation

Materials 

  -digested plasmid backbone
  -equimolar amount digested Part 1
  -equimolar amount digested Part 2
  -T4 DNA ligase buffer
  -T4 DNA ligase
  -dH2O
  -thermal cycler

Procedure

1. Add 2µL of digested plasmid backbone

2. Add equimolar amount of digested Part 1

3. Add equimolar amount of digested Part 2

4. Add 1µL T4 DNA ligase buffer.

5. Add 0.5µL T4 DNA ligase.

6. Add water to 10µL.

7. Place in pre-programmed thermal cycler at 16°C for 30 minutes then at 80°C for 20 minutes.

Retrieved from "http://2013.igem.org/Team:Glendale_CC_AZ/Protocols/Ligation"